Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes.
IF: 8.713
Cited by: 383


The architecture of normal and diseased tissues strongly influences the development and progression of disease as well as responsiveness and resistance to therapy. We describe a tissue-based cyclic immunofluorescence (t-CyCIF) method for highly multiplexed immuno-fluorescence imaging of formalin-fixed, paraffin-embedded (FFPE) specimens mounted on glass slides, the most widely used specimens for histopathological diagnosis of cancer and other diseases. t-CyCIF generates up to 60-plex images using an iterative process (a cycle) in which conventional low-plex fluorescence images are repeatedly collected from the same sample and then assembled into a high-dimensional representation. t-CyCIF requires no specialized instruments or reagents and is compatible with super-resolution imaging; we demonstrate its application to quantifying signal transduction cascades, tumor antigens and immune markers in diverse tissues and tumors. The simplicity and adaptability of t-CyCIF makes it an effective method for pre-clinical and clinical research and a natural complement to single-cell genomics.


cancer biology
computational biology
multiplexed imaging
single-cell method
systems biology

MeSH terms

Antigens, Neoplasm
Immunologic Factors
Microscopy, Fluorescence
Signal Transduction


Lin, Jia-Ren
Izar, Benjamin
Wang, Shu
Yapp, Clarence
Mei, Shaolin
Shah, Parin M
Santagata, Sandro
Sorger, Peter K

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